Rapid Microdetermination of Hydroxyproline in Biomedical Samples by Flow Injection Analysis Using Cysteine as an Antioxidant

Kazuhide UCHIDA; Masako TOMODA; Tetsuichi SHIBATA; Shinobu IKEUCHI; Tetsunori HASEBE; Takeshi MIWA; Tanekuni NOMOTO; Kazuaki FUKUSHIMA; Shin-ichi SAITO; Seiichi INAYAMA

doi:10.2116/analsci.6.39

Kazuhide UCHIDA, Masako TOMODA, Tetsuichi SHIBATA, Shinobu IKEUCHI, Tetsunori HASEBE, Takeshi MIWA, Tanekuni NOMOTO, Kazuaki FUKUSHIMA, Shin-ichi SAITO, Seiichi INAYAMA

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Kazuhide UCHIDA
Department of Anesthesiology, National Defence Medical College
Masako TOMODA
General Science Laboratory, Faculty of Science and Engineering, Sophia University
Tetsuichi SHIBATA
pharmaceutical Institute, School of Medicine, Keio University
Shinobu IKEUCHI
Department of Dentistry and Stomato Surgery, School of Medicine, Keio University
Tetsunori HASEBE
6th Department of Internal Medicine, School of Medicine, Tokai University
Takeshi MIWA
6th Department of Internal Medicine, School of Medicine, Tokai University
Tanekuni NOMOTO
Department of Dentistry and Stomato Surgery, School of Medicine, Keio University
Kazuaki FUKUSHIMA
Department of Anesthesiology, National Defence Medical College
Shin-ichi SAITO
General Science Laboratory, Faculty of Science and Engineering, Sophia University
Seiichi INAYAMA
pharmaceutical Institute, School of Medicine, Keio University

Keywords: Microdetermination, hydroxyproline, collagen, connective tissue, cysteine, antioxidant, flow infection analysis, KISO method, KISOFIA method

JOURNAL FREE ACCESS

1990 Volume 6 Issue 1 Pages 39-44

DOI https://doi.org/10.2116/analsci.6.39

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Published: February 10, 1990 Received: July 31, 1989 Available on J-STAGE: June 30, 2006 Accepted: September 27, 1989 Advance online publication: - Revised: -
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Date of correction: June 30, 2006 Reason for correction: - Correction: ABSTRACT Details: Wrong : >A new practical method for the rapid microdetermination of hydroxyproline (Hyp) in biomedical samples, such as tissue and urine, in the presence of a definite amount of cysteine as antioxidant was established by means of the modified KISO method using flow injection analysis. This method uses no organic solvent and requires only 9min for each sample, which can be injected every 3min (about 5h is sufficient for 100 samples). A good linear relationship (r=0.99996) was obtained between the concentration of Hyp (0-80μg/ml) and the absorbance (0-0.514). In biomedical samples (hydrolyzates of rat stomach tissue), the recoveries (94.8-103.7%) and the reproducibility (the mean of RSD; 0.484% for ten kinds of rat tissues) were satisfactory. The correlation between values from this method and from the conventional KISO method was shown to be good enough (r=0.966) for hydrolyzate samples (n=95), when a proper amount of cysteine solution was added to both the sample and standard solutions and the concentration of the oxidant was optimized to Hyp contents.
Right : A new practical method for the rapid microdetermination of hydroxyproline (Hyp) in biomedical samples, such as tissue and urine, in the presence of a definite amount of cysteine as antioxidant was established by means of the modified KISO method using flow injection analysis. This method uses no organic solvent and requires only 9min for each sample, which can be injected every 3min (about 5h is sufficient for 100 samples). A good linear relationship (r=0.99996) was obtained between the concentration of Hyp (0-80μg/ml) and the absorbance (0-0.514). In biomedical samples (hydrolyzates of rat stomach tissue), the recoveries (94.8-103.7%) and the reproducibility (the mean of RSD; 0.484% for ten kinds of rat tissues) were satisfactory. The correlation between values from this method and from the conventional KISO method was shown to be good enough (r=0.966) for hydrolyzate samples (n=95), when a proper amount of cysteine solution was added to both the sample and standard solutions and the concentration of the oxidant was optimized to Hyp contents.

Date of correction: June 30, 2006 Reason for correction: - Correction: PDF FILE Details: -

Abstract

A new practical method for the rapid microdetermination of hydroxyproline (Hyp) in biomedical samples, such as tissue and urine, in the presence of a definite amount of cysteine as antioxidant was established by means of the modified KISO method using flow injection analysis. This method uses no organic solvent and requires only 9min for each sample, which can be injected every 3min (about 5h is sufficient for 100 samples). A good linear relationship (r=0.99996) was obtained between the concentration of Hyp (0-80μg/ml) and the absorbance (0-0.514). In biomedical samples (hydrolyzates of rat stomach tissue), the recoveries (94.8-103.7%) and the reproducibility (the mean of RSD; 0.484% for ten kinds of rat tissues) were satisfactory. The correlation between values from this method and from the conventional KISO method was shown to be good enough (r=0.966) for hydrolyzate samples (n=95), when a proper amount of cysteine solution was added to both the sample and standard solutions and the concentration of the oxidant was optimized to Hyp contents.

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